Procedures to isolate and purify calcium activated neutral proteinase (CANP II) from bovine brain have been developed by my collaborator, Dr. Banik. In order to determine the bond specificity of CANP II, enzyme digests of human myelin basic (MBP) were made because the sequence of this protein is known. Major peptides of the CANP digestion of MBP have been purified by HPLC and the amino acid composition of them determined. Dr. John Richert has been using our valuable collection of myelin basic proteins and peptides derived from them for the identification of different T-cell reactive sites of other MBP-reactive T-cell clones from normal and multiple sclerosis patients. Two dominant sites of reactivity in MBP were revealed by proliferative responses of T-cell clones from one multiple sclerosis patient. These dominant sites were narrowed to specific regions of MBP by two synthetic peptides (residues 86-105 and 152-170). One of the synthetic peptides (residues 152-170) caused a cytotoxic T lymphocyte response in 19 clones.